scholarly journals Landscape of Bone Marrow Metastasis in Human Neuroblastoma Unraveled by Transcriptomics and Deep Multiplex Imaging

Cancers ◽  
2021 ◽  
Vol 13 (17) ◽  
pp. 4311
Author(s):  
Daria Lazic ◽  
Florian Kromp ◽  
Fikret Rifatbegovic ◽  
Peter Repiscak ◽  
Michael Kirr ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Single Cell ◽  
Tumor Cells ◽  
Bone Marrow Involvement ◽  
Solid Cancer ◽  
Bone Marrow Niche ◽  
Single Cell Level ◽  
Bone Marrow Metastasis ◽  
Cell Level ◽  
Human Neuroblastoma

While the bone marrow attracts tumor cells in many solid cancers leading to poor outcome in affected patients, comprehensive analyses of bone marrow metastases have not been performed on a single-cell level. We here set out to capture tumor heterogeneity and unravel microenvironmental changes in neuroblastoma, a solid cancer with bone marrow involvement. To this end, we employed a multi-omics data mining approach to define a multiplex imaging panel and developed DeepFLEX, a pipeline for subsequent multiplex image analysis, whereby we constructed a single-cell atlas of over 35,000 disseminated tumor cells (DTCs) and cells of their microenvironment in the metastatic bone marrow niche. Further, we independently profiled the transcriptome of a cohort of 38 patients with and without bone marrow metastasis. Our results revealed vast diversity among DTCs and suggest that FAIM2 can act as a complementary marker to capture DTC heterogeneity. Importantly, we demonstrate that malignant bone marrow infiltration is associated with an inflammatory response and at the same time the presence of immuno-suppressive cell types, most prominently an immature neutrophil/granulocytic myeloid-derived suppressor-like cell type. The presented findings indicate that metastatic tumor cells shape the bone marrow microenvironment, warranting deeper investigations of spatio-temporal dynamics at the single-cell level and their clinical relevance.

scholarly journals Heterogeneity of PIK3CA mutational status at the single cell level in circulating tumor cells from metastatic breast cancer patients

2014 ◽  
Vol 9 (4) ◽  
pp. 749-757 ◽  
Author(s):  
Marta Pestrin ◽  
Francesca Salvianti ◽  
Francesca Galardi ◽  
Francesca De Luca ◽  
Natalie Turner ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Metastatic Breast Cancer ◽  
Single Cell ◽  
Cancer Patients ◽  
Tumor Cells ◽  
Metastatic Breast ◽  
Single Cell Level ◽  
Breast Cancer Patients ◽  
Cell Level ◽  
Mutational Status

scholarly journals Probing the Interaction Forces of Prostate Cancer Cells with Collagen I and Bone Marrow Derived Stem Cells on the Single Cell Level

PLoS ONE ◽  
2013 ◽  
Vol 8 (3) ◽  
pp. e57706 ◽  
Author(s):  
Ediz Sariisik ◽  
Denitsa Docheva ◽  
Daniela Padula ◽  
Cvetan Popov ◽  
Jan Opfer ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Stem Cells ◽  
Bone Marrow ◽  
Single Cell ◽  
Cancer Cells ◽  
Collagen I ◽  
Single Cell Level ◽  
Prostate Cancer Cells ◽  
Cell Level ◽  
Interaction Forces

scholarly journals Combined Molecular Genetic and Cytogenetic Analysis from Single Cells after Isothermal Whole-Genome Amplification

2011 ◽  
Vol 57 (7) ◽  
pp. 1032-1041 ◽  
Author(s):  
Thomas Kroneis ◽  
Jochen B Geigl ◽  
Amin El-Heliebi ◽  
Martina Auer ◽  
Peter Ulz ◽  
...  
Keyword(s):  
Single Cell ◽  
Tumor Cells ◽  
Whole Genome Amplification ◽  
Single Cells ◽  
Molecular Genetic ◽  
Target Cells ◽  
Whole Genome ◽  
Single Cell Level ◽  
Cell Level ◽  
Genome Amplification

BACKGROUND Analysis of chromosomal aberrations or single-gene disorders from rare fetal cells circulating in the blood of pregnant women requires verification of the cells' genomic identity. We have developed a method enabling multiple analyses at the single-cell level that combines verification of the genomic identity of microchimeric cells with molecular genetic and cytogenetic diagnosis. METHODS We used a model system of peripheral blood mononuclear cells spiked with a colon adenocarcinoma cell line and immunofluorescence staining for cytokeratin in combination with DNA staining with the nuclear dye TO-PRO-3 in a preliminary study to define candidate microchimeric (tumor) cells in Cytospin preparations. After laser microdissection, we performed low-volume on-chip isothermal whole-genome amplification (iWGA) of single and pooled cells. RESULTS DNA fingerprint analysis of iWGA aliquots permitted successful identification of all analyzed candidate microchimeric cell preparations (6 samples of pooled cells, 7 samples of single cells). Sequencing of 3 single-nucleotide polymorphisms was successful at the single-cell level for 20 of 32 allelic loci. Metaphase comparative genomic hybridization (mCGH) with iWGA products of single cells showed the gains and losses known to be present in the genomic DNA of the target cells. CONCLUSIONS This method may be instrumental in cell-based noninvasive prenatal diagnosis. Furthermore, the possibility to perform mCGH with amplified DNA from single cells offers a perspective for the analysis of nonmicrochimeric rare cells exhibiting genomic alterations, such as circulating tumor cells.

scholarly journals Single-cell landscape of bone marrow metastases in human neuroblastoma unraveled by deep multiplex imaging

2020 ◽  
Author(s):  
Daria Lazic ◽  
Florian Kromp ◽  
Michael Kirr ◽  
Filip Mivalt ◽  
Fikret Rifatbegovic ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Single Cell ◽  
Single Cells ◽  
Therapy Response ◽  
Disseminated Tumor Cells ◽  
Bone Marrow Niche ◽  
Inhibitory Protein ◽  
Apoptotic Pathway ◽  
Human Neuroblastoma ◽  
Bone Marrow Metastases

ABSTRACTBone marrow commonly serves as a metastatic niche for disseminated tumor cells (DTCs) of solid cancers in patients with unfavorable clinical outcome. Single-cell assessment of bone marrow metastases is essential to decipher the entire spectrum of tumor heterogeneity in these cancers, however, has previously not been performed.Here we used multi-epitope-ligand cartography (MELC) to spatially profile 20 biomarkers and assess morphology in DTCs as well as hematopoietic and mesenchymal cells of eight bone marrow metastases from neuroblastoma patients. We developed DeepFLEX, a single-cell image analysis pipeline for MELC data that combines deep learning-based cell and nucleus segmentation and overcomes frequent challenges of multiplex imaging methods including autofluorescence and unspecific antibody binding.Using DeepFLEX, we built a single-cell atlas of bone marrow metastases comprising more than 35,000 single cells. Comparisons of cell type proportions between samples indicated that microenvironmental changes in the metastatic bone marrow are associated with tumor cell infiltration and therapy response. Hierarchical clustering of DTCs revealed multiple phenotypes with highly diverse expression of markers such as FAIM2, an inhibitory protein in the Fas apoptotic pathway, which we propose as a complementary marker to capture DTC heterogeneity in neuroblastoma.The presented single-cell atlas provides first insights into the heterogeneity of human bone marrow metastases and is an important step towards a deeper understanding of DTCs and their interactions with the bone marrow niche.

Dissecting the Functional Reprogramming of the Microenvironment in Bone Marrow Fibrosis at the Single Cell Level

2019 ◽  
Author(s):  
Nils B. Leimkühler ◽  
Li Ronghui ◽  
Hélène F.E. Gleitz ◽  
Inge A. M. Snoeren ◽  
Stijn N.R. Fuchs ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Single Cell ◽  
Single Cell Level ◽  
Cell Level ◽  
Bone Marrow Fibrosis ◽  
Marrow Fibrosis

scholarly journals Development of an optimal protocol for molecular profiling of tumor cells in pleural effusions at single‐cell level

2021 ◽  
Author(s):  
Ikuko Takeda Nakamura ◽  
Masachika Ikegami ◽  
Nobuhiko Hasegawa ◽  
Takuo Hayashi ◽  
Toshihide Ueno ◽  
...  
Keyword(s):  
Single Cell ◽  
Tumor Cells ◽  
Molecular Profiling ◽  
Single Cell Level ◽  
Pleural Effusions ◽  
Cell Level ◽  
Optimal Protocol

scholarly journals Characteristics and requirements of the interaction between human monocytes and tumor cells on the single cell level

Blood ◽  
1983 ◽  
Vol 61 (2) ◽  
pp. 390-396 ◽  
Author(s):  
MG Golightly ◽  
DG Fischer ◽  
C Ohlander ◽  
HS Koren
Keyword(s):  
Tumor Cell ◽  
Single Cell ◽  
Tumor Cells ◽  
Autologous Serum ◽  
Target Cells ◽  
Single Cell Level ◽  
Human Monocytes ◽  
Blood Monocytes ◽  
Cell Level ◽  
Effector Cells

Abstract Highly purified (97%-99%) and viable (99%) peripheral blood monocytes obtained by EDTA-reversible adherence to autologous-serum-precoated plastic surfaces could rapidly lyse a variety of tumor cells in a 3–4 hr 51Cr release assay. Using these monocytes as effectors, a short-term agarose/conjugate assay was utilized, permitting us to examine the interaction between fresh human monocytes and neoplastic target cells on a single cell level. That the tumor-bound effector cells were indeed monocytes was confirmed by employing the monocyte-specific monoclonal antibody 61D3, which stained 95%-99% of the mononuclear cells bound to conjugated and killed K562 tumor targets. The binding of monocytes to target cells appeared to be temperature dependent and was extremely rapid, reaching a plateau after 5 min at 30 degrees C. Our findings demonstrated for the first time that only a proportion of human blood monocytes can bind to a particular target cell and that only a fraction of the binding cells have the intrinsic potential to kill those neoplastic targets. The proportion of monocytes capable of binding and killing varies between individuals and also depends on the tumor cell used, indicating heterogeneity in the monocyte and tumor cell populations. The highest proportion of monocytes bind to the human erythromyeloid leukemia K562 cell line (13%-50%). The frequency of monocytes capable of killing K562 tumor cells is relatively low (7%- 13%). The system described here should be useful to study the heterogeneity of mononuclear phagocytes and to analyze the molecular basis of the interaction between those effector cells and neoplastic target cells.

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